The primary aim of the research in this laboratory is directed towards an understanding of cell wall biosynthesis on the molecular level. Our research emphasizes the role of the membrane in this assembly process. In directing our goals for the project period, areas of research will include the translocation of precursors, regulation of peptidoglycan synthesis, specificity of glycan synthesis, development of new probes for polymer synthesis, and synthesis of membrane teichoic acid. Six specific projects will be investigated. 1. Purification and mechanistic analysis of phospho-MurNAc-pentapeptide translocase. 2. Membrane events in cell wall biosynthesis; a. Spin-labeled intermediates in peptidoglycan synthesis; b. Fluorescent probe in peptidoglycan synthesis; 3. Positive substituent effects on cross-linked peptidoglycan synthesis in a coupled membrane-wall system; 4. Biosynthesis of spin-labeled peptidoglycan; a. Spin-label assay for lysozyme; 5. Regulation of peptidoglycan synthesis in a coupled membrane-wall system; and 6. Biosynthesis of membrane teichoic acid - incorporation of D-alanine. An investigation of these six projects will contribute to our understanding of cell wall assembly and membrane chemistry that relates to this process. Our present objectives will provide insights into the events, regulations, and translocations that occur in the bacterial membrane for elaboration of this extracellular structure, the cell wall.